JP2001039816A - Inhibitor against formation of crosslink in collagen - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain the subject inhibitor free from a problem regarding safety, capable of inhibiting the formation of crosslink in collagen, manifesting aging- resistant effects by repairing flabbiness of the skin and recovering the firm state of the skin, and useful as a skin preparation for external use, etc., by including a medicinal agent such as a hydrolyzed casein and L-lysin. SOLUTION: This inhibitor is obtained by formulating one or more kinds selected from (A) a hydrolyzed casein, (B) L-lysin and (C) L-arginine. For example, the inhibitor is obtained by using the casein obtained by hydrolyzing a milk casein by using a trypsin and a subtilisin by two step reaction, and substituting the hydrolyzed casein to dialysis to desalt and purify the casein, and the preparation for external use can be obtained by formulating about 0.0001-20 wt.% objective inhibitor expressed in terms of a dried material based on the whole amount, and optionally further formulating a bleaching agent, a humectant an antioxidant, an oily component or the like besides the foregoing component.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a collagen cross-linking inhibitor capable of inhibiting collagen cross-link formation by saccharification reaction or oxidation reaction.

[0002]

2. Description of the Related Art The skin is composed of a stratum corneum, an epidermis layer, a basement membrane and a dermis from the outside, and the dermis is the widest part among them. Collagen fibers, elastic fibers, glycoproteins, and proteoglycans have a heterogeneous structure that spreads three-dimensionally in a complex manner, and each structure is in a gel state holding a liquid phase. Collagen fibers are mainly composed of collagen, of which type I collagen accounts for 80% of the whole.

[0003] The collagen fibers in the skin of older people are:
It is harder than that of young people, making it difficult to extract with various solutions (high salt solution, or acetic acid, and even pepsin treatment). This is presumably because crosslinks are formed in and between collagen molecules. The saccharification reaction and the Maillard reaction (Mauron, Prog. Fd. Nutur. Sci., 5:
5-35, 1981). In the case of proteins with slow turnover in the living body such as collagen and extremely long turnover, the influence of sugar on aging is important due to the high rate of long-term sugar modification in living tissue. Have a meaning.

When fibroblasts are cultured in a collagen gel, the collagen gel contracts. The fiber density of the contracted collagen gel is close to the fiber density in the dermis. Modification of this collagen gel with sugars such as glucose suppresses gel contraction by fibroblasts (Howard, Exp. Ce)
11. Res., 228: 132-137. 1998). This is due to the formation of crosslinks within and between collagen molecules due to saccharification,
This is because it becomes difficult for fibroblasts to draw collagen fibers.

In addition to the saccharification reaction, intra- and inter-molecular cross-links are formed by an oxidation reaction, and as a result, cell activity is affected (Tiollier, Exp. Cell. Res., 191: 9).
5-104. 1990).

Therefore, a collagen cross-linking inhibitor which can prevent aging of the skin by inhibiting the formation of collagen cross-links due to saccharification reaction, oxidation reaction and the like and has no problem in terms of safety has been desired.

[0007]

Means for Solving the Problems In order to solve these problems, the present inventors examined the effect of various substances on the inhibition of collagen cross-link formation using collagen gel contraction activity of human skin-derived fibroblasts as an index. As a result, they found that a specific drug has an excellent action, and completed the present invention.

[0008] That is, the present invention is a collagen cross-linking inhibitor comprising one or more selected from the following drugs (1) to (3). (1) Hydrolyzed casein (2) L-lysine (3) L-arginine

The drugs used in the present invention are those already known as general skin external preparations, cosmetics, raw materials for pharmaceuticals, bases and additives. However, the effect of the sugar to inhibit the formation of collagen crosslinks is not known at all.

Hereinafter, the configuration of the present invention will be described in detail. (1) Hydrolyzed casein is obtained by subjecting milk casein to two-stage hydrolysis with trypsin and subtilisin and desalting and purifying it by dialysis. Commercially available products include, for example, Promois Milk (Seiwa Kasei Co., Ltd.). Company-made product name).

As (2) L-lysine and (3) L-arginine, commercially available ones can be used.

The compounding amount of the drug in the present invention is 0.0001 to 20.0% by weight, preferably 0.01 to 5% by weight, as a dry matter, in the total amount of the external preparation. If the amount is less than 0.0001% by weight, the effect of the present invention is not sufficiently exhibited,
If it exceeds 20.0% by weight, it is not preferable because preparation of the composition is difficult.

The drug used in the present invention can achieve the purpose of inhibiting the formation of collagen cross-links and achieving an anti-aging effect by restoring the skin's tallness and firmness.

The collagen crosslinking formation inhibitor of the present invention includes:
In addition to the above essential components, components usually used in external preparations for skin such as cosmetics and pharmaceuticals, for example, whitening agents, humectants, antioxidants, oily components, ultraviolet absorbers, surfactants, thickeners, alcohols, powders Ingredients, coloring materials, aqueous components, water, various skin nutrients, and the like can be appropriately added as necessary.

In addition, sequestering agents such as disodium edetate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, gluconic acid, caffeine, tannin, verapamil, tranexamic acid and derivatives thereof, and licorice extract Products, glabridine, hot water extract of karin fruit, various crude drugs, drugs such as tocopherol acetate, glycyrrhizic acid and derivatives or salts thereof, vitamin C, magnesium ascorbate phosphate, ascorbic acid glucoside, arbutin, kojic acid and the like Other whitening agents, sugars such as glucose, fructose, mannose, sucrose, trehalose and the like can also be appropriately compounded.

The present invention can be widely applied to cosmetics and quasi-drugs applied to the outer skin, particularly preferably cosmetics, and its dosage form can be aqueous, solubilizing, or emulsifying. A wide range of dosage forms can be used, such as powder, oil-liquid, gel, ointment, aerosol, water-oil two-layer systems, water-oil-powder three-layer systems, and the like. That is, if it is a basic cosmetic, face wash, lotion,
It can be widely applied to various forms such as emulsions, creams, gels, essences (packs, masks), etc. In addition, if it is makeup cosmetics,
As a toiletry product such as a foundation, it can be widely applied to forms such as body soap and soap. Furthermore, quasi-drugs can be widely applied to various ointments and the like. And in these dosage forms and forms,
The form that the collagen cross-linking inhibitor of the present invention can take is not limited.

[0017]

The present invention will be described in more detail with reference to the following examples. Note that the present invention is not limited to this. The compounding amount is% by weight. Prior to the examples, an effect test method and its results will be described.

(1) Test Example 1 (Evaluation of the inhibitory effect of cross-link formation by saccharification reaction using collagen gel contraction activity by human dermal fibroblasts as an index) Collagen solution (collagen is IA product, Koken Co., Ltd.)
C was used. ) Was prepared on ice, and added to a 12-well plate for cell culture (Corning: 25815).
The collagen was gelled at 7 ° C. A glucose-6-phosphate solution (final concentration: 10%) containing a test substance (1,3-butylene glycol as a control, the concentration is% by weight)
(0 mM), and the mixture was incubated at 37 ° C. for 7 days to perform a saccharification reaction. Unreacted glucose
After 6-phosphate was removed by washing, 1 × 10 ⁵ cell / ml fibroblasts were inoculated on collagen gel, and 0.25%
The FBS / DMEM medium was loaded. After 3 hours, the gel was peeled off from the petri dish wall, and collagen contraction was performed. 48
After a time, the medium was aspirated, the diameter of the collagen gel was measured, and the area was calculated. Table 1 shows the results. here,
Promoted milk (trade name, manufactured by Seiwa Chemical Co., Ltd.) was used as the hydrolyzed casein.

[0019]

[Table 1] グ リ Glycation Test substance Collagen area after contraction ( cm ² ) ─────────────────────────────────── No Control 247.8 Yes Control 379.9 Yes Hydrolyzed casein (0.01 %) 290.2 Yes Hydrolyzed casein (0.02%) 268.5 Yes Hydrolyzed casein (0.05%) 241.5 Yes Hydrolyzed casein (0.25%) 242.3 Yes Hydrolyzed casein (0.5%) 241.5 Yes L-Lysine (0.01%) 293.3 Yes L -Lysine (0.02%) 255.2 Yes L-Lysine (0.05%) 240.0 Yes L-Lysine (0.25%) 243.2 Yes L-Lysine (0.5%) 243.0 Yes L-Arginine (0.01%) 288.8 Yes L-Arginine (0.02%) ) 266.2 Yes L-arginine (0.05%) 245.5 Yes L-arginine Emissions (0.25%) 240.1 Yes L- arginine (0.5%) 241.2 ───────────────────────────────────

Example 1 Cream (Formulation) Stearic acid 2.0% by weight Stearyl alcohol 7.0 Hydrogenated lanolin 2.0 Squalane 5.0 2-octyldodecyl alcohol 6.0 Polyoxyethylene (25 mol) Cetyl alcohol ether 3.0 Glycerin monostearate 2.0 Propylene glycol 5.0 Hydrolyzed casein (promois milk) 5.0 Tranexamic acid 0.2 Ethyl paraben 0.3 Perfume Appropriate amount Ion exchange water Residue (Production method) To ion exchange water Add propylene glycol,
Heat and maintain at 70 ° C. (aqueous phase). The other components are mixed, heated and melted and kept at 70 ° C. (oil phase). The oil phase is added to the water phase, pre-emulsified, homogenized with a homomixer, and cooled to 30 ° C. with good stirring.

Example 2 Cream (Formulation) Solid paraffin 5.0% by weight Beeswax 10.0 Vaseline 15.0 Liquid paraffin 41.0 Glycerin monostearate 2.0 Polyoxyethylene (20 mol) Sorbitan monolaurate 2 0.0 Soap powder 0.1 Hydrolyzed casein (promois milk) 0.05 Sodium bisulfite 0.03 Ethyl paraben 0.3 Appropriate amount Ion-exchanged water Residue (Preparation method) Add soap powder to ion-exchanged water and heat. 7
Keep at 0 ° C. (aqueous phase). Mix other ingredients, heat and melt.
Keep at 0 ° C. (oil phase). The oil phase is gradually added to the aqueous phase while stirring to carry out the reaction. After completion of the reaction, the mixture is uniformly emulsified with a homomixer, and cooled to 30 ° C. while stirring well after emulsification.

Example 3 Emulsion (Formulation) 2.5% by weight of stearic acid Cetyl alcohol 1.5 Vaseline 5.0 Liquid paraffin 10.0 Polyoxyethylene (10 mol) Monooleate 2.0 Polyethylene glycol 1500 0 Triethanolamine 1.0 Carboxyvinyl polymer 0.05 (Product name: Carbopol 941, BFGoodrich Chemical company) Hydrolyzed casein (promois milk) 0.01 Sodium bisulfite 0.01 Ethyl paraben 0.3 Fragrance Appropriate ion Exchange water residue (Preparation method) Dissolve the carboxyvinyl polymer in a small amount of ion exchange water (phase A). Polyethylene glycol 1500 and triethanolamine are added to the remaining ion-exchanged water, dissolved by heating, and maintained at 70 ° C. (aqueous phase). The other components are mixed, melted by heating and kept at 70 ° C. (oil phase). The oil phase is added to the water phase, pre-emulsification is performed, the phase A is added, and the mixture is uniformly emulsified by a homomixer.

Example 4 Emulsion (Formulation) Microcrystalline wax 1.0% by weight Beeswax 2.0 Lanolin 20.0 Liquid paraffin 10.0 Squalane 5.0 Sorbitan sesquioleate 4.0 Polyoxyethylene (20 mol) Sorbitan monooleate 1.0 Propylene glycol 7.0 Hydrolyzed casein (promois milk) 10.0 Tranexamic acid 1.0 Sodium bisulfite 0.01 Ethyl paraben 0.3 Perfume Appropriate amount Ion-exchange water Residue (Preparation method) Ion Add propylene glycol to the replacement water,
Heat and maintain at 70 ° C. (aqueous phase). The other components are mixed, heated and melted and kept at 70 ° C. (oil phase). While stirring the oil phase, the aqueous phase is gradually added thereto, and the mixture is uniformly emulsified with a homomixer. After emulsification, cool to 30 ° C. while stirring well.

Example 5 Jelly (Formulation) 95% ethyl alcohol 10.0% by weight Dipropylene glycol 15.0 Polyoxyethylene (50 mol) Oleyl alcohol ether 2.0 Carboxyvinyl polymer 0.05 (Product name: Carbopol) 940, BFGoodrich Chemical company) Caustic soda 0.15 L-arginine 0.1 Hydrolyzed casein (promois milk) 0.001 Sodium bisulfite 0.01 Ethyl paraben 0.3 Perfume Appropriate amount Ion exchange water Residue (Production method) Ion exchange water In one step, hydrolyzed casein and polyoxyethylene (50 mol) oleyl alcohol ether are dissolved in 95% ethanol and added to the aqueous phase. Next, after adding other components, the mixture is neutralized with caustic soda and L-arginine to increase the viscosity.

Example 6 Jelly (Formulation) (Phase A) Ethyl alcohol (95%) 10.0% by weight Polyoxyethylene (20 mol) octyldodecanol 1.0 Pantothenyl ethyl ether 0.1 L-lysine 5 Methylparaben 0.15 (B phase) Potassium hydroxide 0.1 (C phase) Glycerin 5.0 Dipropylene glycol 10.0 Sodium bisulfite 0.03 Carboxyvinyl polymer 0.2 (Product name: Carbopol 940, BFGoodrich Chemical company) Purified water Residue (Preparation method) Dissolve A phase and C phase uniformly, and add A to C phase
Add phases and solubilize. Next, after adding the phase B, filling is performed.

Example 7 Pack (Formulation) (A phase) Dipropylene glycol 5.0% by weight Polyoxyethylene (60 mol) hydrogenated castor oil 5.0 (Phase B) L-arginine 0.01 Olive oil 5.0 Acetic acid Tocopherol 0.2 Ethylparaben 0.2 Fragrance 0.2 (Phase C) Hata sodium sulfite water 0.03 Polyvinyl alcohol (degree of saponification 90, degree of polymerization 2,000) 13.0 Ethanol 7.0 Purified water Residue (Production method) A Phase, phase B and phase C are uniformly dissolved,
Add phase B to phase and solubilize. Next, this is added to the C phase and then filled.

Example 8 Sunburn Prevention Cosmetic Stearic Acid 1.5% by Weight Cetyl Alcohol 3.0 Beeswax 2.0 Polyoxyethylene (10 mole addition) Monooleate 1.0 Glycerin Monostearate 1.0 Dimethyl Polysiloxane 10.0 Decamethylcyclopentasiloxane 20.0 2-Hydroxy-4-methoxybenzophenone 3.0 Octyl p-methoxycinnamate 2.0 L-Arginine 0.1 Perfume Appropriate amount Caustic potash Appropriate amount Ion-exchanged water Residue (Production method) After ion-exchanged water and the oil layer components other than the alkali were dissolved by heating (７０70 ° C.), the mixture was neutralized with an alkali, and then emulsified by adding ion-exchanged water. After the emulsification, the mixture was cooled while stirring well.

Example 9 Makeup Base A W / O emulsion type makeup base was prepared with the following composition. (a) Organically modified montmorillonite 0.5% by weight (b) Cetyl isooctanoate 2.0 (c) Octamethylcyclotetrasiloxane 2.0 (d) Decamethylcyclopentasiloxane 5.0 (e) Dimethylpolysiloxane ( 6cs) 5.0 (f) liquid paraffin 3.0 (g) dioctadecyldimethylammonium chloride 0.2 (h) polyoxyalkylene-modified organopolysiloxane 5.0 (i) 4-t-butyl-4'-methoxy Dibenzoylmethane 0.3 (j) Glyceryl mono-2-ethylhexanoyl diparamethoxycinnamate 1.0 (k) Fine particle titanium oxide 5.0 (l) Oleyl alcohol 0.5 (m) Stearic acid 0.5 (n) Sorbitan diisostearate 4.0 (o) Suitable amount of antioxidant (P) Suitable amount of fragrance (q) Talc 1.5 (r) Nylon powder 1.0 (s) Ion exchange water Residue (t) Citric acid Na Lium 0.5 (u) 1,3-butylene glycol 5.0 (v) L-lysine 0.01 (production method) (1) (b)-(j), (l)-(p) and (v) Is heated and dissolved. (2) Add (a) to (1) and disperse and swell. (3) Disperse (k), (q), and (r) in (2). (4) Dissolve (t) and (u) in (s), add to (3) and emulsify.

Example 10 Powdery Foundation (a) Fine Particle Titanium Oxide 7.0% by Weight (b) Talc 40.0 (c) Mica Residue (d) Nylon Powder 10.0 (e) Iron Oxide Red 1.0 (f ) Iron oxide yellow 2.0 (g) Iron oxide black 0.2 (h) Dimethyl polysiloxane 1.0 (i) 2-Ethylhexyl palmitate 9.0 (j) Sorbitan sesquioleate 1.0 (k) N, N-Dimethyl PABA octyl ester 0.3 (l) L-lysine 5.0 (m) Preservative proper amount (n) Antioxidant proper amount (o) Flavor proper amount (Production method) (1) (a)-(g ) Is mixed and pulverized. (2) (h) to (o) are dissolved by heating. (3) After mixing (1) and (2), mold.

Example 11 Oily Foundation (a) Fine Particle Titanium Oxide 10.0% by Weight (b) Mica 22.4 (c) Kaolin 10.0 (d) Nylon Powder 5.0 (e) Iron Oxide Red 0.5 (f) Iron oxide yellow 2.0 (g) Iron oxide black 0.1 (h) Liquid paraffin residue (i) Dimethylpolysiloxane 10.0 (j) Sorbitan sesquioleate 2.0 (k) Octyl methoxycinnamate 5.0 (l) L-arginine 0.005 (m) Perfume appropriate amount (n) Microcrystalline wax 6.0 (o) Carnauba wax 3.0 (Production method) (1) Mix and grind (a) to (g) I do. (2) (h) to (m) are dissolved by heating, and (1) is mixed and dispersed. (3) Grind the slurry of (2) using a grinder. (4) Put (n) and (o) in (3), dissolve by heating, mix and mold.

[0031]

As described above, the collagen cross-linking inhibitor of the present invention has a collagen cross-linking inhibitory activity based on the collagen gel contraction activity of fibroblasts.

 ──────────────────────────────────────────────────続 き Continued on the front page (72) Inventor Shinji Inomata 1050 Nippa-cho, Kohoku-ku, Yokohama, Kanagawa Prefecture Inside Shiseido Daiichi Research Center Co., Ltd. Address F-term in Shiseido Daiichi Research Center Co., Ltd. (reference) 4C083 AA082 AA122 AB032 AB232 AB242 AB352 AB432 AB442 AC012 AC022 AC072 AC082 AC092 AC102 AC122 AC182 AC212 AC242 AC302 AC312 AC352 AC402 AC422 AC432 AC442 AC472 AC482 AC542 AC58 AC582 AC622 AD092 AD152 AD162 AD172 AD421 AD422 AD512 AD662 CC03 CC05 CC07 CC12 CC19 DD31 DD32 DD42 EE12 4C084 AA02 BA43 MA02 MA63 NA14 ZA911 ZC801 4C206 AA01 AA02 FA53 MA03 MA83 ZA91 ZC80

Claims (1)

[Claims] 1. A collagen crosslinking formation inhibitor comprising one or more selected from the following drugs (1) to (3). (1) Hydrolyzed casein (2) L-lysine (3) L-arginine